Development of novel spectroscopic methods for laser-induced dynamic structural change of protein molecules

开发激光诱导蛋白质分子动态结构变化的新型光谱方法

• 批准号: 15204036
• 负责人: KANEMATSU Yasuo
• 金额: $ 31.28万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15204036/
• 关键词: protein; dynamics; laser spectroscopy; fluorescence; infrared light; terahertz spectroscopy; protein crystal; photoreceptor protein; レーザー分光; 光受容蛋白

In order to develop methods for the laser-induced dynamic structural change of protein molecules, we performed the experimental studies of pump-dump-fluorescence measurements and the observation of IR irradiation effect. PYP (photoactive protein) was selected as a target sample protein because it showed an optical cycle leading to its structural change by the irradiation of a triggering light pulse. Furthermore, THz spectroscopy for protein crystals and the synthesis and evaluation of model compounds for PYP were also performed.Pump-dump-fluorescence measurementsPYP exhibits an optical reaction which is observed as the optical cycle with structural change induced by triggering light. In order to investigate the initial stage of the reaction, pump-dump-fluorescence spectroscopy was employed and compared to time resolved fluorescence spectroscopy which enabled us to trace the excited state dynamics through spontaneous emission of light. In the dump process, the excited state population i … More s dumped by stimulated emission process and the dump efficiency was expected to be proportional to the spontaneous emission. However, the detailed evaluation including that for the saturation effect of the dump intensity clarified that the enhancement of the dump efficiency in the longer wavelength regions existed. This result suggested the potential for controllability of the dump process by controlling the pump-and dump pulse light wave.Observation of IR irradiation effectIR light pulse irradiation effect on the optical cycle of PYP was examined for the IR wavelength region (3μ to 6μ). The femtoseconds IR pulses which were created by a hand-made optical parametric amplifier were applied while Nanoseconds light pulses generated by optical parametric oscillator were used for triggering. The 6 μ IR light pulses was observed to enhance the optical reaction in PYP. Although coincident irradiation of the triggering pulses and the IR pulses were observed to enhance the effect, in order to obtain clear evidence, experimental improvements are continuing for quantifying the effect.THz spectroscopyWe constructed a reflection-type THz spectroscopic system and applied to observe the low-frequency modes of protein crystals of lysozyme. The THz spectroscopy for a protein crystal was succeeded for the first time. Observed spectral shapes depending on the humidity of the crystal were in accordance with the results obtained by Raman scattering spectroscopy. Less

为了建立激光诱导蛋白质分子动态结构变化的方法，我们进行了泵浦-倾倒-荧光测量和红外辐射效应观察的实验研究。之所以选择PYP（光活性蛋白）作为靶样蛋白，是因为它在触发光脉冲的照射下表现出光循环导致其结构发生变化。此外，还进行了蛋白质晶体的太赫兹光谱分析和PYP模型化合物的合成和评价。泵浦-转储-荧光测量spyp表现出一种光学反应，其表现为触发光引起结构变化的光循环。为了研究反应的初始阶段，我们采用了泵浦-倾倒-荧光光谱，并将其与时间分辨荧光光谱进行了比较，使我们能够通过自发发光来跟踪激发态动力学。在排土场过程中，受激排放过程所倾倒的激发态粒子数大于受激排放，排土场效率与自发排放成正比。然而，包括倾倒强度饱和效应在内的详细评价表明，在较长波长区域存在倾倒效率的增强。这一结果表明，通过控制泵浦和倾倒脉冲光波，可以实现倾倒过程的可控性。在红外波长范围（3μ ~ 6μ）内考察了有效r光脉冲辐照对PYP光循环的影响。采用手工制作的光参量放大器产生飞秒级红外脉冲，光参量振荡器产生纳秒级红外脉冲进行触发。6 μ IR光脉冲增强了PYP的光学反应。虽然观察到触发脉冲和红外脉冲的同步照射可以增强这种效应，但为了获得明确的证据，实验方面仍在继续改进，以量化这种效应。太赫兹光谱我们构建了一个反射型太赫兹光谱系统，并应用于观察溶菌酶蛋白晶体的低频模式。第一次成功地对蛋白质晶体进行了太赫兹光谱分析。观察到的随晶体湿度变化的光谱形状与拉曼散射光谱的结果一致。少

项目成果

Low-lying singlet states of carotenoids having 8-13 conjugated double bonds as determined by electronic absorption spectroscopy

• DOI: 10.1016/j.cplett.2005.05.037
• 发表时间: 2005-07
• 期刊: Chemical Physics Letters
• 影响因子: 2.8
• 作者: Peng Wang;R. Nakamura;Y. Kanematsu;Y. Koyama;H. Nagae;Tomohiro Nishio;H. Hashimoto;Jianping Zhang-Jianpi

Catena-poly[[[triaquabis(2,6-diacetamidobenzoato)terbium(III)]-m-2.6-diacetamidobenzoato] monohydrate]

链式聚[[[三水双(2,6-二乙酰氨基苯甲酸)铽(III)]-m-2,6-二乙酰氨基苯甲酸]一水合物]

• 发表时间: 2004
• 期刊: Acta Cryst E60
• 作者: A.Onoda;T.Okamura;H.Yamamoto;N.Ueyama
• 通讯作者: N.Ueyama

Photo-induced protein dynamics measured by femtosecond time-resolved luminescence

通过飞秒时间分辨发光测量光诱导蛋白质动力学

• 发表时间: 2003
• 期刊: J. Luminescence Vol.102-103
• 作者: R.Nakamura;Y.Kanematsu;M.Kumauchi;N.Hamada;F.Tokunaga
• 通讯作者: F.Tokunaga

Dynamical process of exciton–exciton scattering in CuI thin films

• DOI: 10.1016/j.jlumin.2006.01.151
• 发表时间: 2007
• 期刊: Journal of Luminescence
• 影响因子: 3.6
• 作者: H. Ichida;T. Shimomura;K. Mizoguchi;Daegwi Kim;Y. Kanematsu;M. Nakayama

O-Atom-Transfer Oxidation of [Molybdenum(IV) Oxo{3,6-(acylamino)2-1,2-benzene dithiolate}2]2- Promoted by Intramolecular NH-S Hydrogen Bonds

分子内NH-S氢键促进的[钼(IV)氧代{3,6-(酰氨基)2-1,2-苯二硫醇}2]2-的O原子转移氧化

• 发表时间: 2006
• 期刊: Inorg. Chem. Vol.45
• 作者: K.Baba;T.Okamura;C.Suzuki;H.Yamamoto;T.Yamamoto;M.Ohama;N.Ueyama
• 通讯作者: N.Ueyama